Large-scale production of extracellular vesicles: Report on the "massivEVs" ISEV workshop.

Extracellular vesicles (EVs) large-scale production is a crucial point for the translation of EVs from discovery to application of EV-based products. In October 2021, the International Society for Extracellular Vesicles (ISEV), along with support by the FET-OPEN projects, "The Extracellular Vesicle Foundry" (evFOUNDRY) and "Extracellular vesicles from a natural source for tailor-made nanomaterials" (VES4US), organized a workshop entitled "massivEVs" to discuss the potential challenges for translation of EV-based products. This report gives an overview of the topics discussed during "massivEVs", the most important points raised, and the points of consensus reached after discussion among academia and industry representatives.

Outer Membrane Vesicles of Actinobacillus pleuropneumoniae Exert Immunomodulatory Effects on Porcine Alveolar Macrophages.

Outer membrane vesicles (OMVs) are spontaneously released by Gram-negative bacteria, including Actinobacillus pleuropneumoniae, which causes contagious pleuropneumonia in pigs and leads to considerable economic losses in the swine industry worldwide. A. pleuropneumoniae OMVs have previously been demonstrated to contain Apx toxins and proteases, as well as antigenic proteins.

Clarification of large-volume bacterial cultures using a centrifuge-free protocol.

Aims: To provide a reliable, reproducible and centrifuge-free filtration protocol for clarification of large volumes of bacterial cultures.

Methods And Results: Four experiments were designed to compare different techniques enabling clarification of Escherichia coli cultures using as a benchmark the concentration and quality of bacterial outer membrane vesicles (OMVs). The experiments were designed to examine the performance of different extraction methods on large volume (≥1 L) filtrations of bacterial culture media.

Estimating upper-extremity function from kinematics in stroke patients following goal-oriented computer-based training.

Introduction: After a stroke, a wide range of deficits can occur with varying onset latencies. As a result, assessing impairment and recovery are enormous challenges in neurorehabilitation. Although several clinical scales are generally accepted, they are time-consuming, show high inter-rater variability, have low ecological validity, and are vulnerable to biases introduced by compensatory movements and action modifications.

Bacterial Outer Membrane Vesicles as a Versatile Tool in Vaccine Research and the Fight against Antimicrobial Resistance.

Gram-negative bacteria include a number of pathogens that cause disease in humans and animals. Although antibiotics are still effective in treating a considerable range of infections caused by Gram-negative bacteria, the alarming increase of antimicrobial resistance (AMR) induced by excessive use of antibiotics has raised global concerns. Therefore, alternative strategies must be developed to prevent and treat bacterial infections and prevent the advent of a postantibiotic era.

Exact results on high-dimensional linear regression via statistical physics.

It is clear that conventional statistical inference protocols need to be revised to deal correctly with the high-dimensional data that are now common. Most recent studies aimed at achieving this revision rely on powerful approximation techniques that call for rigorous results against which they can be tested. In this context, the simplest case of high-dimensional linear regression has acquired significant new relevance and attention.

Demonstration of Self-Starting Nonlinear Mode Locking in Random Lasers.

In ultrafast multimode lasers, mode locking is implemented by means of saturable absorbers or modulators, allowing for very short pulses. This occurs because of nonlinear interactions of modes with well equispaced frequencies. Though theory predicts that, in the absence of any device, mode locking would occur in random lasers, this has never been demonstrated so far.

Effect of Detergents on Morphology, Size Distribution, and Concentration of Copolymer-Based Polymersomes.

Polymersomes made of amphiphilic diblock copolymers are generally regarded as having higher physical and chemical stability than liposomes composed of phospholipids. This enhanced stability arises from the higher molecular weight of polymer constituents. Despite their increased stability, polymer bilayers are solubilized by detergents in a similar manner to lipid bilayers.

Design and Characterization of a Novel Tool for the Antigenic Enrichment of Outer Membrane.

Production and isolation of recombinant proteins are costly and work-intensive processes, especially in immunology when tens or hundreds of potential immunogens need to be purified for testing. Here we propose an alternative method for fast screening of immunogen candidates, based on genetic engineering of recombinant bacterial strains able to express and expose selected antigens on their outer membrane. In , a Gram-negative porcine pathogen responsible for extensive economic losses worldwide, we identified a conserved general secretion pathway (GSP) domain in the N-terminal part of the outer membrane protein ApfA (ApfA stem: ApfA).

Exploiting phage receptor binding proteins to enable endolysins to kill Gram-negative bacteria.

Bacteriophage-encoded endolysins degrading the bacterial peptidoglycan are promising antibacterials for combating antibiotic-resistant bacteria. However, endolysins have limited use against Gram-negative bacteria, since the outer membrane prevents access to the peptidoglycan. Here, we present Innolysins, an innovative concept for engineering endolysins to exert antibacterial activity against Gram-negative bacteria.

Hydrostatic Filtration Enables Large-Scale Production of Outer Membrane Vesicles That Effectively Protect Chickens against .

is a Gram-negative opportunistic avian pathogen representing an emerging threat to poultry meat and egg production worldwide. To date, no vaccine able to effectively prevent the morbidity associated with infections has been developed yet. Our group previously reported that inoculation of different combinations of outer membrane vesicles (OMVs), FlfA and GtxA-N proteins is effective in preventing lesions caused by infections in layer chickens.

Immunoproteomic characterization of outer membrane vesicles from hyper-vesiculating Actinobacillus pleuropneumoniae.

Outer membrane vesicles (OMVs) are produced and secreted virtually by every known Gram-negative bacterium. Despite their non-live nature, they share antigenic characteristics with the bacteria they originate from. This, together with their relative ease of purification, casts the OMVs as a very promising and flexible tool in both human and veterinary vaccinology.

The membrane transporter PotE is required for virulence in avian pathogenic Escherichia coli (APEC).

Over the last few years, polyamines have been described as key-signal of virulence in pathogenic bacteria. In the current study, we investigated whether the knockout of genes related to polyamine biosynthesis and putrescine transport affected the virulence of an avian pathogenic E. coli (APEC) strain.

In vivo testing of novel vaccine prototypes against Actinobacillus pleuropneumoniae.

Actinobacillus pleuropneumoniae (A. pleuropneumoniae) is a Gram-negative bacterium that represents the main cause of porcine pleuropneumonia in pigs, causing significant economic losses to the livestock industry worldwide. A.

Identification and characterization of serovar-independent immunogens in Actinobacillus pleuropneumoniae.

Despite numerous actions to prevent disease, Actinobacillus pleuropneumoniae (A. pleuropneumoniae) remains a major cause of porcine pleuropneumonia, resulting in economic losses to the swine industry worldwide. In this paper, we describe the utilization of a reverse vaccinology approach for the selection and in vitro testing of serovar-independent A.

Inverse problem for multi-body interaction of nonlinear waves.

The inverse problem is studied in multi-body systems with nonlinear dynamics representing, e.g., phase-locked wave systems, standard multimode and random lasers.

The glassy random laser: replica symmetry breaking in the intensity fluctuations of emission spectra.

The behavior of a newly introduced overlap parameter, measuring the correlation between intensity fluctuations of waves in random media, is analyzed in different physical regimes, with varying amount of disorder and non-linearity. This order parameter allows to identify the laser transition in random media and describes its possible glassy nature in terms of emission spectra data, the only data so far accessible in random laser measurements. The theoretical analysis is performed in terms of the complex spherical spin-glass model, a statistical mechanical model describing the onset and the behavior of random lasers in open cavities.

General phase diagram of multimodal ordered and disordered lasers in closed and open cavities.

We present a unified approach to the theory of multimodal laser cavities including a variable amount of structural disorder. A general mean-field theory is studied for waves in media with variable nonlinearity and randomness. Phase diagrams are reported in terms of optical power, degree of disorder, and degree of nonlinearity, tuning between closed and open cavity scenarios.

Small-cluster renormalization group in Ising and Blume-Emery-Griffiths models with ferromagnetic, antiferromagnetic, and quenched disordered magnetic interactions.

The Ising and Blume-Emery-Griffiths (BEG) models' critical behavior is analyzed in two dimensions and three dimensions by means of a renormalization group scheme on small clusters made of a few lattice cells. Different kinds of cells are proposed for both ordered and disordered model cases. In particular, cells preserving a possible antiferromagnetic ordering under renormalization allow for the determination of the Néel critical point and its scaling indices.

[Oral premedication with clonidine as an alternative in dental practice. The effects on the pain threshold, blood pressure and salivary flow].

Background: Clonidine, an alpha 2-adrenoceptor agonist, has been recently shown to be effective in conscious sedation because of its analgesic and sedative properties, having also proven to be active in reducing neuroendocrine responses to stressful stimuli perioperatively. The current study was designed to investigate the efficacy of 150 micrograms oral clonidine as a premedicant in dentistry.

Methods: In a prospective, randomized, double-blind controlled clinical trial, 40 patients, aged 16-64 yr, undergoing conservative, or prosthetic, or dental surgery procedures, received placebo (n = 20) or 150 micrograms of oral clonidine (n = 20) 90 min before the estimated time of induction of local anesthesia.

[Papilloma of the palate. Clinical and histopathology].

An asymptomatic, exophytic and unusually large mass of hard and soft palate is presented. Its papillary surface architecture is characteristic of an oral squamous papilloma. The cause is unknown.

[Severe tooth agenesis. Presentation of a case].

In the present work the Authors describe a case of a patient affected by Duchenne disease as well as by dental multiple agenesis. It is a particular case of such rare syndromes in the same subject.

[Subcutaneous emphysema during intraligamental anesthesia].

Periodontal ligament anaesthesia (PDLA) is a good choice to most common local anaesthesia methods. In the present work the Authors report a case of subcutaneous emphysema as adverse effect of this injection technique.